Enhancement of homology-directed repair with chromatin donor templates in cells
Enhancement of homology-directed repair with chromatin donor templates in cells
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DOI：
10.7554/elife.55780
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发表时间：
2020-04-28
期刊：
影响因子：
7.7
通讯作者：
Kadonaga, James T.
中科院分区：
文献类型：
--
作者：
Cruz-Becerra, Grisel;Kadonaga, James T.
精确基因组编辑的一个关键挑战是同源定向修复（HDR）的低效率。在这里，我们描述了一种通过使用染色质供体模板而不是裸 DNA 供体模板来提高细胞 HDR 效率的策略。染色质（细胞核中 DNA 的天然形式）的使用增加了 HDR 编辑克隆以及纯合编辑的频率。此外，染色质转染导致的细胞毒性可以忽略不计。这些发现表明，染色质供体模板应该可用于广泛的 HDR 应用，例如精确插入或替换包含基因编码区的 DNA 片段。
A key challenge in precise genome editing is the low efficiency of homology-directed repair (HDR). Here we describe a strategy for increasing the efficiency of HDR in cells by using a chromatin donor template instead of a naked DNA donor template. The use of chromatin, which is the natural form of DNA in the nucleus, increases the frequency of HDR-edited clones as well as homozygous editing. In addition, transfection of chromatin results in negligible cytotoxicity. These findings suggest that a chromatin donor template should be useful for a wide range of HDR applications such as the precise insertion or replacement of DNA fragments that contain the coding regions of genes.